Analysis of the main factors involved in the evaluation of Prunus resistance to Plum pox virus (Sharka) in controlled greenhouse conditions

In countries like Spain or France, quarantine rules force researchers to evaluate the resistance to Sharka (Plum pox virus, PPV) in controlled, isolated conditions. This evaluation method shows important limitations resulting from the management of plants in the controlled conditions, grown in pots with artificial cycles of growth in the greenhouse and cold chamber, alternately. The objective of this study is to analyse different factors that affect the efficiency of the method of evaluation of PPV resistance in controlled greenhouse conditions. The cultivars evaluated as model genotypes were the resistant ‘Stark Early Orange’ and the susceptible ‘Real Fino’ apricot. Furthermore the ‘GF305’ peach was used as a susceptible control. The different studied factors were the inoculation protocol (rootstock or variety inoculation), the grafting success (depending on inoculation method, rootstock–variety combination and date of grafting), and the efficiency of the process in each artificial cycle of growth. Results showed that rootstock inoculation was more effective than inoculation of the variety. As rootstock, the ‘GF305’ seedlings were slightly better than the ‘Real Fino’ seedlings in the inoculation process, but they were quite similar in terms of effectiveness in the evaluation and grafting process. Grafting can be carried out in spring or autumn without having important differences. The global efficiency of the evaluation process was much higher with rootstock inoculation.     

Soil and rootstock influence on fruit quality of plums (Prunus domestica L.)

Soil and rootstock can particularly affect the ‘Rainha Claudia Verde’ (Prunus domestica) fruit quality, mainly its firmness characteristics. To investigate the variation in fruit quality, plums were harvested at commercial maturity from trees grafted on Marianna GF8-1 (Prunus cerasifera × munsoniana) and Marianna GF10-2 (P. cerasifera × munsoniana) rootstocks on two different soils: Haplic Luvisol and Vertic Luvisol. After harvest fruits were stored at 2 °C for 3 weeks. At the harvest day a small group of fruits were analysed without cold storage. During storage fruits were tested for firmness, soluble solids content, titratable acidity and fruit mineral content. Tree vigour was evaluated from trunk cross-sectional areas values.The GF8-1 rootstock promoted the highest vegetative development, comparing to GF10-2. This rootstock promoted the largest fruits size and the higher calcium fruit level. Positive correlations were found between higher concentrations of calcium in the pulp fruits and firmness. Fruits from two different rootstocks showed the same firmness at harvest but during cold storage, fruits from GF10-2 rootstock exhibited the highest firmness pulp values. There weren’t significant differences in fruit nitrogen and potassium levels for any studied factor.At harvest solids soluble content and solids soluble content/titratable acidity ratio weren’t affected by soil type or rootstock. As expected, solids soluble content during cold storage increased and solids soluble content/titratable acidity had a slight increase.GF10-2 revealed to be a good option as a ‘Rainha Claudia Verde’ rootstock for plums growers due to its intermediate-vigour. Also GF10-2 fruits presented better quality during storage comparing with GF8-1.     

Physiological alterations modulated by rootstock and scion combination in cashew under salinity

In this study we evaluated the influence of rootstocks and scions on physiological disturbances that are induced by salinity in cashew (Anacardium occidentale) plantlets. Two cashew genotypes, CCP 09 and BRS 226, were utilized as rootstocks and scions, resulting in four scion/rootstock combinations by reciprocal grafting. The plantlets were irrigated in absence (control) or in presence of 50 or 200 mM NaCl for 15 days under greenhouse conditions. The plantlets with BRS 226 as rootstocks demonstrated higher transpiration and greater accumulations of Na⁺ and Cl⁻, proline and free amino acids in leaves compared to plantlets that having CCP 09 as rootstocks. The K⁺ content in roots and leaves of all four combinations was not influenced by salinity or by different scion/rootstock combinations. The self-grafting of the BRS 226 genotype showed the highest stability for chlorophyll and Rubisco, exhibiting the highest tolerance to salinity. The scion genotype did not affect any of the studied physiological parameters. The studied physiological disturbances induced by salinity in cashew plantlets were more influenced by rootstock than by scion and these changes were also dependent on compatibility between scion and rootstock.     

Characterization for yield, fruit quality, and molecular profiles of lemon genotypes tolerant to ‘mal secco’ disease

‘Mal secco’ is a serious disease of lemon and other citrus species, reduces lemon production, and limits the use of susceptible cultivars especially in the Mediterranean countries, against which there is no effective management system to control the disease. The objective of this study was to determine yield efficiency, tree and fruit characters, and molecular profiles of the selected promising genotypes in lemon breeding program to develop ‘mal secco’ tolerant lemon cultivars. After 4 years of evaluation, ‘Tuzcu 894’, a progeny of ‘Finike Yerli Yuvarlak’ and ‘Kutdiken’, was designated as promising genotype with high yield, fruit weight, juice content and low seed number contrary to ‘Kutdiken’ as control. ‘Tuzcu 8911’ was suitable for dense planting having the highest yield in CV. In addition, these two promising genotypes survived after many years being subjected to artificial and natural P. tracheiphila infections and harsh winter temperatures, which were attractive to the citrus industry. Molecular analysis, as assessed with 11 inter-simple sequence repeats (ISSR) primers, indicated that ‘Tuzcu 09 Aklimon’ and its selections, ‘Tuzcu 896’, ‘Tuzcu 897’ and ‘Tuzcu 898’, were distinct from the other genotypes with similarity value of 0.82. Genetic variation among the other lemon genotypes was low and some of the genotypes were identical. It was concluded that variations in the agronomical characters are mainly due to mutations.     

Evaluation of apricot resistance to Plum pox virus (Sharka) in controlled greenhouse and natural field conditions

In this study we compare the evaluation of Plum pox virus (Sharka) resistance of 29 apricot genotypes in controlled greenhouse conditions by grafting onto infected ‘GF305’ peach seedlings growing in pots, and in natural conditions by grafting onto infected 5-year-old apricot trees growing in the orchard. Apricot genotypes evaluated were initially classified into three groups: susceptible to PPV (presence of PPV symptoms and ELISA positive in greenhouse and field assays), resistant (absence of PPV symptoms and ELISA negative in both assays) and undetermined (evaluated differently in both assays). Results showed a similar behavior against Plum pox virus in both assays in 20 out of the 29 apricot genotypes studied (69%). However, in the other nine genotypes results were different (31%). Evaluation in field was more accurate, detecting a higher number of susceptible genotypes, probably due to the higher inoculation pressure in the old trees in comparison with the GF305 rootstocks in pots. However, greenhouse evaluation let to work in controlled isolated conditions with a higher number of genotypes. This situation is greatly required in areas where Sharka is not widely spread. Greenhouse evaluation could be then the first screening method to evaluate Plum pox virus resistance of apricot genotypes, and could be complemented with the evaluation onto infected trees in natural conditions in insect-proof quarantine shelter.     

Production in Vitro of Rootstock/Scion Combinations of Prunus Cultivars

Shoot meristems (0-2-0-5 mm) of the plum scion cv Victoria (Prunus domestica) and the plum rootstock cv Pixy (P. insititia) were micrografted in vitro onto rooted shoots of the peach rootstock GF305 (P. pérsica) produced by in vitro propagation. These procedures, which resulted in 40—50% successful grafts, are preliminaries to providing a more controlled system for research into rootstock influence on scion growth and development.     

Fruit quality parameters of ‘Pioneer’ Japanese plums produced on eight different rootstocks

The influence of Marianna GF 8-1, GF 677, Julior-Ferdor, Nemared, Monegro (G × N 9), Felinem (G × N 22), Adesoto 101 and Cadaman rootstocks on fruit quality of ‘Pioneer’ Japanese plum was analysed. The study was performed during three consecutive years in the Guadalquivir Valley (Seville, Spain), on a silt loam and calcareous soil. Colour, fruit size, fruit and stone weights, hardness, soluble solid concentration and acidity were measured for fruit quality evaluation. In general, the effect of the different rootstocks on most of the analysed quality parameters was variable because a strong interaction rootstock × year was observed. However, several parameters as fruit shape, soluble solid concentration, acidity and maturity seem to be harder conditioned in some rootstocks, did not show significant differences year-by-year. Fruits produced on scions grafted on Marianna and Nemared showed the highest soluble solid concentration and Cadaman the lowest, the last also originated the hardest fruits and the lowest maturity.     

Transformation of Brassica oleracea var. capitata with bacterial betA gene enhances tolerance to salt stress

The bacterial betA gene for biosynthesis of glycinebetaine was transferred to cabbage (Brassica oleracea var. capitata) cultivar ‘Golden Acre’ through Agrobacterium-mediated transformation of hypocotyl explants. Transgenic status was established through Southern hybridization and mRNA expression in the shoots. The transformants exhibited higher tolerance to NaCl stress compared to untransformed parent plants. In physiological assessment of salinity tolerance, transgenics showed better growth response and greater stability in maintaining plant water relations at increasing levels of salinity. These results demonstrate that engineering glycinebetaine biosynthetic pathway into cabbage can lead to enhanced salt tolerance.     

Identification of host plant resistance to pepper leaf curl virus in chilli (Capsicum species)

Three hundred and seven genotypes belonging to four cultivated and one wild species of Capsicum were screened against pepper leaf curl virus (PepLCV) causing devastating leaf curl disease of chilli (Capsicum annuum). Initial screening was done under field conditions based on coefficient of infection (CI), disease reaction to each genotype was assigned. Subsequently, selfed progenies of eight symptom-less and highly resistant lines were challenged by viruliferous white fly under glasshouse conditions, out of which only three genotypes, viz. GKC-29, BS-35 and EC-497636 showed no symptom. Using scion and root stalk of susceptible genotype (Pusa Jwala), these three putative symptom-less genotypes were further challenged by grafting and alternate grafting. The resistant reactions of GKC-29, BS-35, EC-497636 were confirmed because even after 50 days of successful grafting/alternate grafting, no viral symptom appeared on all the grafted plants of these genotypes. When subjected to PCR amplification with degenerate primers deigned to detect gemnivirus like PepLCV, the three symptom-less genotypes did not show any amplification, suggesting that the resistant reaction in three identified symptom-less resistant sources was because of the absence of viral genome and they are not symptom-less carrier.     

Immunolocalization and quantification of IAA after self- and free-pollination in Olea europaea L.

In order to characterise the self-compatibility in ‘Pisa 9’, that is an improved clone of cultivar Leccino (Olea europaea L.), biological, histological and biochemical analyses were carried out. Observations were also focused on ‘Pendolino’, the main pollinator for Italian cultivars considered as being self-incompatible. After self-pollination, in ‘Pisa 9’ the majority of the pollen tubes reached the distal part of the style, entered the ovule leading to a high fruit-set percentage. These same processes were also noted in the Pendolino cultivar, though in a lesser degree. The indole acetic acid (IAA) concentration and its immunocytochemical localization were determined in pollinated flowers after self- and free-pollination and in unpollinated flowers. In both cultivars the highest IAA levels and signal intensity were observed in free-pollinated pistils, whereas after self-pollination a different behaviour emerged. In ‘Pendolino’ the immunostaining was not uniformly distributed and stopped at the style level. This finding sustained the low percentage of fruit-set obtained in the cultivar. In ‘Pisa 9’ the auxin distribution was similar to the free-pollinated pistils where the immunolocalization staining reached the ovary. The correspondence between the IAA signal in the ovary and the high percentage of fruit-set in ‘Pisa 9’ self-pollinated pistils, confirms the role of this hormone in the induction and regulation of the fertilisation process.     

Flavonol and phenolic acid composition of sweet cherries (cv. Lapins) produced on six different vegetative rootstocks

The effect of rootstock (‘MaxMa 14’, ‘Weiroot 13’, ‘PiKu 1’, ‘Weiroot 158’, ‘Gisela 5’ and ‘F12/1’) on phenolic acid and flavonol content of “Lapins” sweet cherry was investigated. Phenolic acids and flavonols were isolated from sweet cherries and analyzed by using reversed phase high-performance liquid chromatography (RP-HPLC). The major phenolic acids in sweet cherries were neochlorogenic acid (18–50 mg kg⁻¹), chlorogenic acid (19–62 mg kg⁻¹) and p-coumaric acid derivatives (15–125 mg kg⁻¹). The amount of flavonol quercetin-3-rutinoside (8–37 mg kg⁻¹) was significant as well. There are significant variations in the phenolic compound content among sweet cherry fruits grown on trees grafted on different vegetative rootstocks. The significantly higher chlorogenic acid, neochlorogenic acid, p-coumaric derivative and quercetin-3-rutinoside contents were found in sweet cherry fruits grown on trees grafted on ‘Weiroot 13’ and ‘PiKu 1’ rootstocks. Sweet cherries produced on trees grafted on other rootstocks had significantly lower phenolic compound content.     

Effect of explants source on shoot proliferation and adventitious regeneration in 10 Buddleia cultivars

Viable shoot cultures and weaned plants were obtained from cultured apical meristems with 10 Buddleia cultivars giving viabilities of 32–72%. The number of shoots produced, the micropropagation rate and the root number produced in vitro was higher in meristem derived shoots compared to those derived from shoot-tips. The subsequent growth rate of meristem derived plants, in the greenhouse, was also greater. The number of roots produced by conventional cuttings collected from meristem derived plants was significantly higher than in cuttings which were collected from plants derived from shoot-tips or from the original stock plants.Endogenous bacteria were not detected in either shoot cultures derived from meristems or in 10-week-old weaned plants derived from meristems whereas those derived from shoot-tips showed the presence of endogenous bacteria when sterilized explants were cultured on nutrient agar or on tryptic soy broth.Factors affecting adventitious bud and shoot production in leaf and internode explants was determined for ‘Lochinch’, ‘Border Beauty’, ‘Ile de France’ and ‘Pink Delight’ using meristem derived shoot cultures. Adventitious shoots appeared after 4 weeks of culture, in both types of explant when cultured on MS supplemented with 0.5–5.0 μM TDZ. The highest percentage regeneration was achieved from bisected internode explants cultured on 0.5 μM TDZ, with 93–100% regeneration among the cultivars whereas BA was less effective. The best response was obtained using 5.0 μM TDZ which gave over 10–11 shoots per explant in all bisected explants for all cultivars.     

Auxins increase the occurrence of leaf-colour variants in Caladium regenerated from leaf explants

Leaf explants of Caladium ‘Pink Cloud’ were cultured in vitro on MS medium containing various auxins (NAA, IBA, IAA, 2,4,5-T and 2,4-D) in combination with cytokinin (BA). NAA gave the most vigorous in vitro propagation of this plant, and only 15% of the plants were leaf-colour variants on the medium containing 0.5 μmol NAA. Leaf colour variation was observed in all plants regenerated on the medium containing 2,4-D at 0.5–4.5 μmol. In hormone-free medium, only a few leaf-colour variants (6%) occurred, but the rate of plant regeneration was very low. Application of 0.5 μmol NAA together with 4.5 μmol BA seemed to be the most appropriate for in vitro propagation of Caladium ‘Pink Cloud’ with only a few leaf-colour variants.     

Development of a male sterile eggplant by utilizing the cytoplasm of Solanum virginianum and a biparental transmission of chloroplast DNA in backcrossing

To develop a male sterile line of eggplant (Solanum melongena L.) utilizing the cytoplasm of S. virginianum, an interspecific F₁ hybrid between S. virginianum and S. melongena ‘Senryo Nigou’ was continuously backcrossed to S. melongena ‘Uttara’ using ‘Uttara’ as a recurrent pollen parent and four backcross generations, BC₁, BC₂, BC₃ and BC₄ were produced. All the plants in backcross generations were anther indehiscent although the F₁ hybrid, S. virginianum and S. melongena were dehiscent. Pollen stainability with acetic carmine and in vitro germination ability of pollen in all the backcross progenies were quite lower than those of the parental species. Fruit set percentage, number of seeds per fruit and seed germination rate were high in all the backcross progenies. The present results indicate that anther indehiscent type of functional male sterile line of eggplant could be developed by utilizing the cytoplasm of S. virginianum. PCR-RFLP analysis of chloroplast DNA (cpDNA) and mitochondrial DNA (mtDNA) were performed to identify the organelle inheritance. The F₁ hybrid and all the backcross progenies displayed maternal inheritance of mtDNA. The cpDNA of the examined single BC₁ plant exhibited the recombinant cpDNA pattern of the parental F₁ hybrid and ‘Uttara’, indicating occurrence of the biparental inheritance of cpDNA. As all the BC₂, BC₃ and BC₄ progenies showed the same recombinant cpDNA patterns of the BC₁ plant, the recombinant cpDNA might be stable and harmonize with the nuclear genome of S. melongena. The present male sterile line can contribute to expand the male sterility source of eggplant.     

Morphogenic gradients of adventitious bud and shoot regeneration in epicotyl explants of Citrus

The in vitro responses of epicotyl explants from ‘Cravo’ rangpur lime (Citrus limonia Osb.), ‘Foster’ grapefruit (C. paradisi Macf.), and ‘Pera’ sweet orange (C. sinensis (L.) Osb.) were characterized for the first time. Further analysis was performed in ‘Cravo’ rangpur lime and ‘Foster’ grapefruit aiming to verify the in vitro morphogenesis of five distinct regions of the epicotyl under different treatments. It was observed the same general pattern of morphogenic gradient along the epicotyl axis in both citrus cultivars, with greater organogenic response as the distance of the explants from the cotyledonary node increased. This morphogenic gradient was influenced by factors related to plant material, composition of the culture medium, and conditions of incubation. The regions of the epicotyl farthest from cotyledons could be used as a source of explants in experiments of genetic transformation of the genotypes evaluated aiming to improve the efficiency of production of transgenic Citrus plants.     

Chill unit models for blackcurrant (Ribes nigrum L.) cultivars ‘Ben Gairn’, ‘Ben Hope’ and ‘Ben Tirran’

Temperate-zone crops require a period of winter chilling to terminate dormancy and ensure adequate bud break the following spring. The exact chilling requirement of blackcurrant (Ribes nigrum), a commercially important crop in northern Europe, is relatively unknown. Chill unit models have been successfully utilized to determine the optimum chilling temperature of a range of crops, with one chill unit equating to 1 h exposure to the optimum temperature for chill satisfaction. Two-year-old R. nigrum plants of the cultivars ‘Ben Gairn’, ‘Ben Hope’ and ‘Ben Tirran’ were exposed to temperatures of −10.1 °C, −3.4 °C, 0.1 °C, 1.5 °C, 2.1 °C, 3.4 °C or 8.9 °C (±0.7 °C) for durations of 0, 2, 4, 6, 8 or 10 weeks and multiple regression analyses used to determine the optimum temperature for chill satisfaction.     

Field performance and molecular diversification of lemon selections

Lemon (Citrus limon (L.) Burm. f.) is one of the most important Citrus fruit for Turkey because of its great amount of production and export. It has been cultivated for a long time in Turkey, and therefore variations for agronomical traits are likely among cultivated lemons due to bud mutations and, hybridizations. The objectives of this study were to determine variations for some selected agronomical traits and genetic markers among 12 new lemons derived from selections. Tree growth, yield, fruit quality, and molecular diversification of these clones were determined. After four years of evaluation, ‘Kutdiken’ M-51 indicated the highest canopy volume. For yield per tree, the best clone was ‘Kutdiken’ M-51. After five years of evaluation, ‘Kibris’ M-54 had the highest fruit weight and acidity. ‘Italian Memeli’ M-56 contained the lowest seed number and the highest total soluble solids. Molecular analysis, as assessed with 22 random amplification of polymorphic DNA (RAPD) and 11 inter simple sequence repeats (ISSR) primers, indicated that seven of twelve clones were separated with RAPD markers, whereas four were distinguished with ISSR markers. Combined analysis of RAPD and ISSR data detected that similarity values among the lemons clones were between 0.97 and 1.00. It can be concluded that variations in orchards are abundant and mainly due to mutations.     

An assessment of the drought tolerance of Fraxinus genotypes for urban landscape plantings

The Fraxinus genus consists of ca. 65 deciduous, rarely evergreen trees found mainly throughout Europe, Asia and North America. Fraxinus species possess the aesthetic characteristics desired of urban trees (autumn colour, attractive bark and flowers); however, no scientifically based tolerance ranking exists for many genotypes within this genera. Aims of this study were to identify whether differences in drought tolerance exist within the Fraxinus genus and thereby provide information as to their usefulness for planting in urban landscapes where water stress poses a threat to transplant survival. At the cessation of a 2-week drought stress and subsequent recovery period, marked differences in drought tolerance among genotypes were distinguished based on net photosynthesis, leaf chlorophyll fluorescence and chlorophyll concentration. In addition, measurement of light absorbance, light trapping, electron transport and dissipation fluxes per leaf cross section of photosystem II at the cessation of the drought period provided an insight into mechanisms of drought damage and resilience among the genotypes tested. The results of this investigation indicate that F. excelsior ‘Aurea Pendula’, F. nigra Marsh., F. ornus L., F. angustifolia ‘Raywood’, F. excelsior L. and F. excelsior ‘Jaspidea’ should be considered for urban planting sites, where water supply is limited. Care should be taken when planting F. americana L., F. americana ‘Autumn Purple’ and F. velutina Torr., into urban environments as results indicated these species are sensitive to drought.     

Eggplant relatives as sources of variation for developing new rootstocks: Effects of grafting on eggplant yield and fruit apparent quality and composition

We propose the utilization of eggplant (Solanum melongena L.) interspecific hybrids derived from crosses with closely related species as an approach for developing new improved rootstocks for eggplant. Here we investigate rootstock effects on fruit yield, apparent quality and proximate and mineral composition of S. melongena ‘Black Beauty’ (BB) scions grafted on interspecific hybrid rootstocks developed from crosses of S. melongena with Solanum incanum L. (SI × SM) and Solanum aethiopicum L. (SM × SA). The results are compared with non-grafted (BB control) and self-grafted (BB/BB) controls and with S. melongena ‘Black Beauty’ scions grafted onto Solanum torvum Sw. (STO) and Solanum macrocarpon L. (SMA) rootstocks. All treatments were grown in a soil naturally infested with root-knot nematodes (mostly Meloidogyne incognita (Kofoid and White) Chitwood). SI × SM and SM × SA interspecific hybrids had high germination (≥90%) and total graft success (100%). Contrary to what occurred with all other treatments, no plants from scions grafted onto these hybrid rootstocks died during the experiment. In particular, the SI × SM hybrid rootstock conferred the highest vigour to the scion, which resulted in the highest values for fruit earliness and early and total yield. Little difference was observed among treatments for apparent fruit quality traits, except for a greater fruit calyx length and prickliness of fruit grafted onto SMA rootstocks. A similar result was obtained for fruit composition where phenolics content was higher in fruit from plants grafted onto SMA rootstocks. Grafting eggplant onto interspecific eggplant hybrids, especially on the SI × SM hybrid, has proved advantageous for eggplant production, as the high vigour and good compatibility of the rootstock with scion results in improved early and total yield without negative effects on apparent fruit quality or composition. Interspecific hybrids represent an alternative to the commonly used STO rootstock, which is a wild species with irregular germination.